Developing Methanosarcina spp. as a model system to study cytochromes c and their role in archaeal methane metabolism

正在开发甲烷八叠球菌属。

基本信息

批准号：
10679362
负责人：
Dinesh Gupta
金额：
$ 6.95万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-30 至 2025-09-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10679362
关键词：
Anaerobic Bacteria Archaea Atmosphere Binding Biochemical Biogenesis Biological Models Biotechnology CRISPR/Cas technology Coculture Techniques Complex Consumption Coupled Development Electron Transport Electrons Engineering Environment Escherichia coli Evaluation Ferredoxin Gene Expression Genes Genetic Genetic Models Genome Genomics Goals Growth Health Heme Human In Vitro Knowledge Mediating Membrane Metabolic Methane Methane Metabolism Pathway Methanobacteria Methanosarcina Methanosarcina barkeri Modeling Molecular NADH Nature Nitrogen Fixation Operon Organism Oxidation-Reduction Pathway interactions Physiological Play Postdoctoral Fellow Process Production Proteins Reaction Research Rhodobacter Role Source Symbiosis System Techniques biochemical tools citrate carrier climate change climate crisis cofactor cytochrome c genome editing greenhouse gases improved in vivo insight interest methanophenazine microorganism oxidation reconstitution sodium ion success sulfate reducing bacteria technology development tool

项目摘要

Project Summary/Abstract Cytochromes c are crucial in methane-metabolizing archaea for the production and consumption of methane coupled to growth and energy conservation. Overall, methane-metabolizing archaea mediate the net flux of methane released into the atmosphere and thus, significantly impact the global methane cycle and climate change. Based on genomic studies, Cytochromes c have been hypothesized to play an important role in methane metabolism however the underlying molecular mechanisms remain elusive, primarily due to the absence of a well-developed genetic model system. Even attempts to study archaeal cytochrome c using an alternative approach such as heterologous expression in well-established bacterial systems have not proven successful so far. The main focus of the proposed research is to develop a genetically tractable methanogenic archaeon, Methanosarcina acetivorans, as a platform to functionally characterize archaeal cytochrome c and gain physiological insights into the role of cytochrome c in methane metabolism across different archaeal species. Cytochromes c are ubiquitous electron transfer proteins that require a covalent attachment to its heme co-factor, a process called cytochrome c biogenesis. Coordination between the cytochrome c biogenesis pathway and the cytochrome c of interest is critical for the successful production of a functional cytochrome c in a heterologous host. Using genetic and biochemical tools, I have recently characterized the cytochrome c biogenesis pathway in the model methanogenic archaeon, Methanosarcina acetivorans. Using this knowledge, the project aims to develop M. acetivorans as a genetic chassis to produce and functionally characterize archaeal cytochrome c from diverse archaeal species. Aim1 of the project will functionally characterize the crucial cytochromes c belonging to methane-producing archaea or methanogens, and Aim2 will study these proteins from methane-consuming archaea using both in vitro and in vivo analyses. This research will improve our understanding of methane metabolism and lead to the development of an archaeal host to study cytochrome c proteins from archaea. Ultimately, the knowledge garnered from these studies can be used to develop sustainable solutions for the global climate crisis and mitigate its harmful impacts on human health.

项目概要/摘要细胞色素 c 在甲烷代谢古菌中对甲烷的生产和消耗至关重要与增长和能量守恒相结合。总体而言，甲烷代谢古菌介导了甲烷的净通量甲烷释放到大气中，从而对全球甲烷循环和气候产生重大影响改变。基于基因组研究，细胞色素 c 被假设在然而，甲烷代谢的潜在分子机制仍然难以捉摸，主要是由于缺乏完善的遗传模型系统。甚至尝试使用替代方法，例如在成熟的细菌系统中进行异源表达尚未得到证实到目前为止成功。拟议研究的主要重点是开发一种基因可控的产甲烷菌古细菌，Methanosarcina acetivorans，作为功能表征古细菌细胞色素 c 和获得关于细胞色素 c 在不同古菌甲烷代谢中的作用的生理学见解物种。细胞色素 c 是普遍存在的电子转移蛋白，需要与其血红素辅因子共价连接，称为细胞色素 C 生物发生的过程。细胞色素 c 生物发生途径与感兴趣的细胞色素 c 对于在异源细胞中成功生产功能性细胞色素 c 至关重要主持人。使用遗传和生化工具，我最近描述了细胞色素 c 生物发生途径在产甲烷古菌模型中，Methanosarcina acetivorans。利用这些知识，该项目旨在开发 M. acetivorans 作为遗传底盘来生产古细菌细胞色素 c 并对其进行功能表征来自不同的古菌物种。该项目的目标 1 将在功能上描述关键的特征细胞色素 c 属于产甲烷古菌或产甲烷菌，Aim2 将研究这些蛋白质使用体外和体内分析从消耗甲烷的古细菌中提取。这项研究将改善我们的了解甲烷代谢并导致开发出用于研究细胞色素 c 的古菌宿主来自古细菌的蛋白质。最终，从这些研究中获得的知识可用于开发全球气候危机的可持续解决方案并减轻其对人类健康的有害影响。